Reliable reference genes for gene expression analyses under the hypomagnetic field in a migratory insect.
Reliable reference genes for gene expression analyses under the hypomagnetic field in a migratory insect.
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用于迁徙昆虫低磁场下基因表达分析的可靠参考基因
DOI：
10.3389/fphys.2022.954228
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发表时间：
2022
影响因子：
4
通讯作者：
中科院分区：
文献类型：
--
作者：
操纵低磁场（HMF），即地磁场（GMF）的缺失或显著减弱（<5 μT），为研究磁场对生物生理，发育，行为和生活史的影响提供了独特的工具。逆转录定量聚合酶链反应（RT-qPCR）已用于研究与暴露于HMF相关的基因表达变化。然而，选择合适的参考基因（RG）与RT-qPCR的环境中确认稳定表达往往是低估。使用三种算法（BestKeeper，Normandy和GeNorm），我们研究了8个候选RG的表达稳定性时，暴露于HMF条件与当地的GMF从青少年到成年的迁移性昆虫害虫，褐飞虱褐飞虱发育过程中。在若虫期，RPL 5 & α-TUB 1、EF 1-α & ARF 1、RPL 5 & AK、EF 1-α & RPL 5和ARF 1 & AK分别是最稳定的RG组。对于1 ~ 3日龄成虫，AK & ARF 1、AK & α-TUB 1、AK & ARF 1和EF 1-α & RPL 5、AK & α-TUB 1、AK & EF 1-α分别是长翅型和短翅型成虫的最佳RG集合。ACT 1 & RPL 5、RPL 5 & EF 1-α、α-TUB 1 & ACT 1和EF 1-α & RPL 5、ARF 1 & ACT 1、ACT 1 & ARF 1分别是长翅型和短翅型雄虫的最佳RG组。这些结果将有助于准确的基因表达分析下的HMF在N。lugens。在这项研究中所示的验证方法强调了确定可靠的RG磁生物学（包括magnetoreception），涉及磁场强度作为一个因素的未来实证研究的重要性。
Manipulating the hypomagnetic field (HMF), which is the absence or significant weakening (<5 μT) of the geomagnetic field (GMF), offers a unique tool to investigate magnetic field effects on organismal physiology, development, behavior and life history. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) has been utilized to study changes in gene expression associated with exposure to the HMF. However, selecting appropriate reference genes (RGs) with confirmed stable expression across environments for RT-qPCR is often underappreciated. Using three algorithms (BestKeeper, NormFinder, and GeNorm), we investigated the expression stability of eight candidate RGs when exposed to the HMF condition versus local GMF during developmental from juveniles to adults in the migratory insect pest, the brown planthopper Nilaparvata lugens. During the nymphal stage, RPL5 & α-TUB1, EF1-α & ARF1, RPL5 & AK, EF1-α & RPL5, and ARF1 & AK were suggested as the most stable RG sets in the 1st to 5th instars, respectively. For 1- to 3-day-old adults, AK & ARF1, AK & α-TUB1, AK & ARF1 and EF1-α & RPL5, AK & α-TUB1, AK & EF1-α were the optimal RG sets for macropterous and brachypterous females, respectively. ACT1 & RPL5, RPL5 & EF1-α, α-TUB1 & ACT1 and EF1-α & RPL5, ARF1 & ACT1, ACT1 & ARF1 were the optimal RG sets for macropterous and brachypterous males, respectively. These results will facilitate accurate gene expression analyses under the HMF in N. lugens. The verification approach illustrated in this study highlights the importance of identifying reliable RGs for future empirical studies of magnetobiology (including magnetoreception) that involve magnetic field intensity as a factor.
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影响因子：
6
作者：
Binhi VN;Rubin AB
通讯作者：
Rubin AB
电磁场通过哭泣的依赖性途径破坏了果蝇中的负岩石。
影响因子：
16.6
作者：
Fedele, Giorgio;Green, Edward W.;Rosato, Ezio;Kyriacou, Charalambos P.
通讯作者：
Kyriacou, Charalambos P.
影响因子：
4.5
作者：
Fedele G;Edwards MD;Bhutani S;Hares JM;Murbach M;Green EW;Dissel S;Hastings MH;Rosato E;Kyriacou CP
通讯作者：
Kyriacou CP
DOI：
10.1016/j.jphotobiol.2018.05.027
发表时间：
2018-08-01
影响因子：
5.4
作者：
Agliassa, Chiara;Narayana, Ravishankar;Maffei, Massimo E.
通讯作者：
Maffei, Massimo E.
影响因子：
16.6
作者：
Foley, Lauren E.;Gegear, Robert J.;Reppert, Steven M.
通讯作者：
Reppert, Steven M.